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Precipitation of LDL with sulfated polyanions: three methods compared.

Three precipitation methods for the determination of low density lipoproteins have been evaluated. In n = 113 normolipidemic samples mean LDL-cholesterol levels have been 2.90 mmol/l, 2.77 mmol/l, 3.21 mmol/l after precipitation with heparin, dextran sulfate, and polyvinylsulfate, respectively. As compared to a combined ultracentrifugation and precipitation reference procedure (mean 3.22 mmol/l) two precipitation methods tend to underrate LDL-cholesterol. Elevated plasma triglycerides may interfere with the precipitation of LDL. The clinical relevance of the precipitation procedures has been studied by discriminant analysis in n = 28 consecutive patients admitted for coronary bypass operation and n = 28 controls. The data suggest that, statistically, the determination of LDL-cholesterol with either precipitation method only provides redundant information as in relation to the Friedewald approximation for LDL-cholesterol. Immunologically determined apolipoprotein B proved a better predictor for group separation than either precipitation method.

Adult↗

Amniotic fluid phospholipids. Study of the cold acetone precipitation effect.

Amniotic fluid phospholipids were extracted and separated with chloroform: methanol (2:1 by vol). These phospholipids were also precipitated with cold acetone and the percentage of each phospholipid precipitated was calculated. The precipitation percentage of the six phospholipids studied ranged from 33.78 +/- 0.70 (mean +/- SEM) for lecithin to 93.77 +/- 1.40 for phosphatidylserine. The variation of L/S, PG/S and PI/S ratios depending on the precipitation step was also studied. We observed that cold acetone precipitation decreased L/S and PG/S ratios, whereas PI/S remained unchanged. Our results indicated that: (1) cold acetone precipitation affected each phospholipid in a different way; (2) the precipitation step was not an effective way of completely separating surfactant and non-surfactant material; (3) a new predictive value should be established if the L/S ratio without precipitation is to be used in the clinic on a daily basis.

Acetone↗

Interaction of bovine gallbladder mucin and calcium-binding protein: effects on calcium phosphate precipitation.

BACKGROUND & AIMS: Gallstones consist of calcium salts and cholesterol crystals, arrayed on a matrix of gallbladder mucin (GBM), and regulatory proteins like calcium-binding protein (CBP). To determine if interactions between CBP and GBM follow a biomineralization scheme, their mutual binding and effects on CaHPO4 precipitation were studied. METHODS: Binding of CBP to GBM was assessed by inhibition of the fluorescence of the complex of GBM with bis-1,8-anilinonaphthalene sulfonic acid (bis-ANS). The effects of the proteins on precipitation of CaHPO4 were assessed by nephelometry and gravimetry. Precipitates were analyzed for calcium, phosphate, and protein. RESULTS: CBP and bis-ANS competitively displaced each other from 30 binding sites on mucin, with a 1:1 stoichiometry and similar affinity. The rate of precipitation of CaHPO4 was retarded by mucin and CBP. Precipitate mass was unaffected by GBM alone but decreased with the addition of CBP. Complexing CBP with GBM abolished or moderated this latter effect, altered precipitate morphology, and changed the stoichiometric ratios of Ca to PO4 in the precipitates from 1:1 to 3:2. Mucin and CBP were incorporated into the precipitates. CONCLUSIONS: These studies suggest that the formation of calcium-containing gallstones is a biomineralization process regulated by both GBM and CBP.

Anilino Naphthalenesulfonates↗

Composition of IgA immune complexes precipitated with polyethylene glycol. A model for isolation and analysis of immune complexes.

Covalently cross-linked large and intermediate-sized IgA oligomers, prepared with IgA anti-dinitrophenyl (DNP) and bis-DNP-pimelic acid ester, were used to examine the ability of different concentrations (3.5%, 5% and 7%, w/v) of polyethylene glycol (PEG) to precipitate IgA immune complexes (IgA-IC). The size of the IgA-IC precipitated with PEG was determined by gradient polyacrylamide gel electrophoresis and quantitative autoradiography. The standard concentration of 3.5% PEG precipitated only a minor fraction (20%) of the IgA-IC. In contrast, 5% and 7% PEG precipitated 45% and 79% of the complexes, respectively. To test the influence of the antigen on the PEG assay. IgA-IC prepared with IgA anti-DNP and DNP conjugates of either bovine serum albumin or Ficoll were also used. Approximately 38% of these IgA-IC were precipitated with 3.5%, PEG. By comparison, the concentration of 5% and 7% PEG precipitated 60% and 76% of the IgA-IC, respectively. Distilled water rather than the standard borate-buffered saline was shown to be the optimal solvent for resolubilization of the PEG precipitates. Serum samples from 22 IgA nephropathy patients and 12 normal donors were tested with 3.5%, 5% and 7% PEG. Only the 7% PEG assay showed a significant difference between patients and controls (P less than 0.001) in the IgA levels of precipitates. Thus, the use of 7% PEG is recommended for the detection, isolation and analysis of large- and intermediate-sized IgA-IC.

Antigen-Antibody Complex↗

The asymmetric IgG non-precipitating antibody. Localization of the oligosaccharide involved, by concanavalin A interaction.

Binding to Con A-Sepharose 4B of the low-affinity Fab fragment from sheep IgG1 anti-Dnp non-precipitating antibody was previously determined. This communication reports the results obtained when the concanavalin A interaction with Fd' fragments and L chains from non-precipitating and precipitating antibodies was examined. When Fd' fragments and L chains from non-precipitating and precipitating antibodies were tested as inhibitors of the binding of 125I-labelled concanavalin A to guinea pig erythrocytes, inhibition of such binding was only achieved by Fd' fragments from non-precipitating antibodies. Forty eight per cent of this Fd' fragment was bound by Con A-Sepharose 4B. From these results and our previous studies we conclude that mannose and/or glucose residues must be present and exposed at the Fd' fragment from the low-affinity Fab arm of sheep IgG1 anti-Dnp non-precipitating antibody. Glycosylation differences may explain the difference in precipitation behaviour of the 2 types of antibody.

Animals↗

Advantages of sequential polyethylene glycol (PEG) precipitations to isolate and analyze circulating immune complexes.

The present report describes a new approach utilizing sequential polyethylene glycol 6000 (PEG) precipitations to isolate and analyze circulating immune complexes (IC). Two experimental systems were studied. First, IC were formed by adding labeled or unlabeled bovine serum albumin (BSA) to anti-BSA antiserum. Sequential precipitations with PEG concentrations ranging from 2 to 6.2 indicated that immunoglobulin (Ig) molecules were precipitated in all experimental and control sera tested. In contrast, optimal precipitations of IC (i.e., highest BSA/Ig ratio) were a function both of PEG concentration and of BSA/anti-BSA ratio (i.e., of IC size). Satisfactory IC purification was achieved by precipitation with appropriate PEG concentration followed by incubation with STaphylococcus aureus, Cowan I. Second, in preliminary experiments, the same procedure was applied to human sera. The findings paralleled those of the BSA/anti-BSA system. While PEG precipitated Ig in all sera tested, a potential antigen was found only in the most IC-positive serum, and only at the higher of the two PEG concentrations tested. From our data, we conclude that PEG concentrations needed to precipitate IC cannot be predetermined, as they are a function of the size and probably the nature of the IC studied. This limitation can be overcome by the systematic sequential precipitations which increase the chances of identifying antigens and decrease the risks of confusing antigen-antibody complexes and Ig aggregates.

Animals↗

Canine bile contains anticrystallization factors that inhibit precipitation of calcium carbonate.

Previous studies have strongly suggested that human bile contains a substance(s) that interferes with the precipitation of calcium phosphate and carbonate from solution. These studies, however, did not distinguish between calcium binding by biliary constituents resulting in decreased calcium carbonate saturation (alterations in solution thermodynamics) and true inhibition of calcium salt precipitation by kinetic factors. Because our recent studies have shown that canine common duct bile is always supersaturated with calcium carbonate (thermodynamically at risk for precipitation), we hypothesized that it must contain kinetic factors that inhibit formation and/or growth of calcium carbonate crystals. Effects of canine bile, bovine albumin and the bile salt taurocholate on calcium carbonate precipitation were studied in highly supersaturated solutions of calcium carbonate that spontaneously undergo rapid precipitation. Measured free ionized calcium concentrations, [Ca++], and calculated calcium carbonate saturation indices were compared in test solutions and controls to evaluate the thermodynamic effects of test solutions on the degree of saturation in the assay system. It is shown that addition of only 0.2 ml of normal canine gallbladder bile to the assay system (a 1:101 dilution of biliary components) abolished precipitation. A lesser inhibitory effect (a decrease in the rate of precipitation) was observed when gallbladder bile was diluted but was lost after 10-fold dilution. Canine common duct bile caused a decrease in the rate of precipitation similar to diluted gallbladder bile. In contrast, sodium taurocholate (250 mmol/L), the major bile salt in the dog, and albumin (1.5 gm/L), the most abundant protein in bile, had only a minimal inhibitory effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Lysozyme purification from tobacco extract by polyelectrolyte precipitation.

Tobacco is widely used as a model plant for feasibility studies of recombinant protein production from transgenic plants. However, dealing with large quantities of biomass to recover recombinant proteins is a challenge for down-stream processing. In this study, the effect of isoelectric precipitation on native tobacco protein was first studied. Among the three acids studied, hydrochloric acid is shown to be more effective than acetic or citric acid, and at pH 4, 60% of native tobacco protein was precipitated by HCl. Egg white lysozyme was used as the model protein to test the feasibility of polyelectrolyte precipitation in protein recovery from tobacco extract. Precipitation of lysozyme at pH 7 was shown ineffective probably because of the interference of polyphenolic acids. However, after isoelectric precipitation at pH 5 poly(acrylic) acid (PAA) was shown to precipitate 85% of the soluble lysozyme when the polymer dosage was increased to 1.5 mg polymer/mg lysozyme, while negligible amounts of native tobacco protein was co-precipitated. Lysozyme precipitation by PAA in tobacco extract obtained at pH 5 was also studied, and lysozyme yield was significant improved.

Chemical Precipitation↗

Polyethyleneimine precipitation versus anion exchange chromatography in fractionating recombinant beta-glucuronidase from transgenic tobacco extract.

Tobacco has been studied as a possible host for the production of recombinant proteins. In this report, recombinant beta-glucuronidase (rGUS) was used as a model protein to study the feasibility of using polyethyleneimine (PEI) precipitation to fractionate acidic recombinant proteins from transgenic tobacco. Results showed that rGUS was preferentially precipitated when the PEI dosage was beyond 200mg PEI/g total protein. At 700-800 mg PEI/g total protein, nearly 100% rGUS was precipitated with less than 40% native tobacco proteins co-precipitated. Approximately 85-90% of the rGUS activity could be recovered from the precipitation pellet for an enrichment ratio of 2.7-3.4. As a comparison, anion exchange chromatography (AEX) yielded similar results to PEI precipitation with 66-90% rGUS activity recovered and an enrichment ratio of 1.8-3.1. The rGUS was further purified by an additional hydrophobic interaction chromatographic (HIC) step after precipitation or AEX. Similar results in terms of rGUS activity recovered and enrichment were obtained. The major interfering protein at the end of all purification steps is most likely the Fraction 1 protein ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco). The presence of this protein is likely the cause for the varying and somewhat low enrichment ratios, but it may be later removed via a size-exclusion chromatography step. PEI precipitation offers the advantage of allowing significant sample concentration prior to subsequent purification techniques such as chromatography and is much less expensive than chromatographic methods as well. Through direct comparison, this study shows that PEI may be used as an initial fractionation step in replacement of AEX to facilitate the purification of acidic recombinant proteins from transgenic tobacco.

Animals↗

Precipitation scavenging of 7Be and 137Cs radionuclides in air.

Atmospheric depositional fluxes of the naturally occurring 7Be of cosmogenic origin and 137Cs from fallout of the Chernobyl accident were measured over a 6-year period (January 1987-December 1992) at Thessaloniki, Greece (40 degrees 38'N, 22 degrees 58'E). Total precipitation accumulation during 1987-1992 varied between 33.7 cm and 65.2 cm, reflecting a relatively dry (precipitation-free) climate. The activity concentrations of 7Be and 137Cs in rainwater depended on the precipitation rate, being higher for low precipitation rates and lesser for high precipitation rates. 137Cs was removed by rain and snow more efficiently than 7Be. Snowfall was more efficient than rainfall in removing the radionuclides from the atmosphere. The annual bulk depositional fluxes of 7Be varied between 477 and 1133 Bq m(-2) y(-1) and this variability was attributed to the amount of precipitation and the variations of the atmospheric concentrations of 7Be. The annual bulk depositional fluxes of 137Cs showed a significant decrease over time from 1987 to 1992, resulting in a removal half-life of 1.33 years. The presence of 137Cs in air, and therefore in rainwater and snow, long after the Chernobyl accident (26 April 1986) was mainly due to the resuspension process. The normalized depositional fluxes of both radionuclides showed maximal values during the spring season where the maximum amount of precipitation occurred. The relatively high positive correlation between 7Be and 137Cs normalized depositional fluxes indicates that the scavenging process of local precipitation controlled the fluxes of both radionuclides. The dry depositional flux of 7Be was less than 9.37% of total (wet and dry) depositional flux. The fraction of dry-to-total depositional flux of 137Cs was much higher than that of 7Be, due to the resuspended soil.

Air Pollutants, Radioactive↗

The influences on leachate from landfill of incineration residuals by acid precipitation.

Incineration of municipal solid wastes (MSW) is the main method of waste management in Taiwan. Although the incineration of MSW processes the solid wastes at 850-950 degrees C and destroys most of the organics, the content of incineration ashes is still a problem for landfill. Moreover, acid precipitation is much worse than before in Taiwan, especially in the northern areas. For instance, the occurrence probabilities of acid precipitation measured from 1991 to 1998 in Taipei increase from 73% to 85%. Therefore, it is more important to get a series of data that will help explore the influence of acid precipitation during disposal on characterization of pollutants than to analyze the ash properties after the incinerators have been constructed and regularly used. In this investigation, the disposal site of incineration ashes is simulated in laboratory by test columns. An irrigation experiment is taken to simulate the acid precipitation at room temperature. In order to explore the exact influence on leachate quality of the main chemical composition of acid precipitation, columns are migrated with different concentrations of sulfate in acid precipitation. This investigation showed that the sulfate concentration of acid precipitation has an increasing effect on the accumulative release of heavy metals, such as Zn, Pb and Cu, from leachate. The sulfate concentration of acid precipitation, however, will not influence the trend of chemical oxygen demand (COD), biochemical oxygen demand (BOD(5)) and total organic carbon (TOC) in the leachate release.

Acids↗

Contribution of marine and continental aerosols to the content of major ions in the precipitation of the central Mediterranean.

The region of the investigated receptor is situated in the southern part of the Adriatic Sea in the Mediterranean. The measuring station is located on the seashore, which, being considered as a border area, is representative for the qualitative and quantitative estimation of the influence of marine and continental aerosols on the content of major ions in precipitation. In the sampling period, precipitation in the region of the investigated receptor was more abundant during the summer and autumn than during the winter and spring. The most frequent precipitation heights were up to 20 mm, while high precipitation came exclusively from the continental region. The results of the measurements of ions readily soluble in water were used for the differentiation of marine from continental contributions of primary and secondary aerosols to their content in the precipitation. Using PCA, it was shown that main contribution of Cl(-), Na(+) and Mg(2+) came from primary marine aerosols, while the contribution from continental sources was dominant for the content of SO(4)(2-), NO(3)(-), NH(4)(+) and Ca(2+) in the precipitation. The continental origin of Ca(2+) was from a primary source, while SO(4)(2-), NO(3)(-) and NH(4)(+) were representatives of secondary aerosols produced by reactions between acid oxides and alkaline species in the atmosphere, but SO(4)(2-) and NO(3)(-) also exist in the precipitation as free acids. The origin of the trace elements Cd, Cu, Pb and Zn in the precipitation came from anthropogenic emission sources. The results obtained in this work are based on experimental data from 609 samples collected during the period 1995-2000.

Aerosols↗

Modelling multiple mineral precipitation in anaerobic digester liquor.

Mineral precipitation problems have been experienced with the conveyance and treatment of anaerobically digested primary and waste activated sludge blends. This paper describes an experimental investigation into mineral precipitation in anaerobic digester liquor (ADL) from the Cape Flats (CF) Wastewater Treatment Plant (WWTP) (Cape Town, South Africa), and application of the three-phase (aqueous/solid/gas) physical and chemical processes kinetic model developed by Musvoto et al. (Water Res. 34 (2000) 1857; Water Res. 34 (2000) 1868; Water SA 26(4) (2000) 417) to the experimental data. From the experimental investigation and theoretical modelling, it is concluded inter alia that: (i) there is a close correlation between experimental measured and theoretically predicted data, (ii) the dominating mineral that precipitates is struvite, with small amounts of amorphous calcium phosphate and negligible newberyite, calcite and magnesite, (iii) the precipitation of struvite is governed by the increase in pH when CO2 is lost from the ADL, (iv) the ADL is initially undersaturated with respect to struvite, but becomes supersaturated at pH > 7.3-7.7, (v) the rate and mass of struvite precipitation are controlled by the rate of pH increase and the initial Mg concentration and (vi) the three-phase kinetic model is able to simulate accurately the time dependent precipitation data for multiple minerals competing for the same species and allows determination of specific precipitation rates for a number of minerals simultaneously in an integrated manner from a single batch test. Some operational strategies to minimise struvite precipitation are proposed.

Bacteria, Anaerobic↗

Fibrinolytic activity of salivary euglobulin fractions precipitated at pH 5.9 or 6.4 before and after mixing with blood.

Mixed saliva was collected from 20 healthy young persons before oral surgery, immediately after the operation and at control 5 days later. The fibrinolytic activity of untreated whole saliva and salivary euglobulin fractions precipitated at pH 5.9 (10 patients) or at pH 6.4 (10 patients) was measured on human fibrin plates. The salivary euglobulin fractions were also processed on plasminogen-free fibrin plates. The concentrations of alpha 1-antitrypsin, antithrombin III, C1-esterase inhibitor, alpha 2-macroglobulin, alpha 2-plasmin inhibitor and plasminogen were measured in pooled plasma and four postoperative saliva samples before and after preparing the corresponding dissolved euglobulins precipitated at pH 5.9 or pH 6.4. Dissolved euglobulins from the pooled plasma were subjected to crossed immunoelectrophoresis with antisera to plasminogen in the second-dimension gel. Normal mixed saliva showed unaltered fibrinolytic activity after euglobulin precipitation, whereas euglobulins prepared from postoperative samples had higher activity than the untreated specimens. No significant differences of fibrinolytic activity between salivary euglobulins precipitated at pH 5.9 and 6.4 were demonstrated. The recovery of proteinase inhibitors in the euglobulin fractions of plasma is lower than 2.1% except C1-esterase inhibitor, which is precipitated, but only at pH 5.9. The mixing with saliva does not seem to change this relative quantity of precipitated inhibitors from plasma. Plasminogen precipitated from plasma is not proteolytic degraded and the yield is higher at pH 5.9 than at pH 6.4. Salivary euglobulins precipitated at pH 5.9 or 6.4 are superior to untreated saliva to measure the fibrinolytic activity of mixed saliva, especially in samples contaminated with blood.

Blood↗

Isoelectric precipitation of soybean protein using carbon dioxide as a volatile acid.

A novel process is presented for the isoelectric precipitation of soy protein, using carbon dioxide as a volatile acid. By contacting a soy meal extract with pressurized carbon dioxide, the solution pH was decreased to the isoelectric region of the soy proteins. Complete precipitation of the precipitable soy proteins could be achieved for protein concentrations up to 40 g/l at pressures less than 50 bar. Isoelectric precipitation with a volatile acid enabled accurate control of the solution pH by pressure and eliminated the local pH overshoot, usual in conventional precipitation techniques. The advantage of the improved precipitation control was reflected by the morphology of the precipitate particles. Protein aggregates formed by CO2 were perfectly spherical whereas protein precipitated by sulfuric acid had an irregular morphology. The influence of process variables to control particle size is discussed.

Carbon Dioxide↗

Alkalinization and precipitation characteristics of 0.2% ropivacaine.

BACKGROUND AND OBJECTIVES: Alkalinization of local anesthetics has been used to increase the speed of onset of nerve blocks. However, alkalinization of local anesthetic solutions may cause precipitation, thereby decreasing bioavailability and anesthetic activity. Alkalinization of ropivacaine has not been described. This laboratory study assessed the alkalinization and precipitation characteristics of ropivacaine. METHODS: Aliquots (2 mL) of commercially available ropivacaine (Naropin, 0.2%; Astra Pharmaceutical, Westborough, MA) were alkalinized with increasing amounts (0.01, 0.02, 0.04 mL) of sodium bicarbonate (8.4%) and immediately monitored for pH change and onset of visible precipitation at room temperature. We then alkalinized ropivacaine with sodium bicarbonate and measured the amount of precipitate that accumulated after various incubation times. RESULTS: The pH of ropivacaine increases with the addition of small amounts of bicarbonate. The calculated percentage of nonionized ropivacaine increased from 0.3% to greater than 30% with alkalinization from pH of 5.51 to 7.63. Drug loss to precipitation increased with higher doses of bicarbonate, reaching 25% to 30% of the total ropivacaine. Even with a low dose of bicarbonate (0.1 mL bicarbonate/20 mL ropivacaine), precipitation increased with time of incubation, reaching a plateau at 20 minutes. CONCLUSIONS: A laboratory evaluation that establishes the alkalinization characteristics of ropivacaine is a prerequisite for designing a clinical study of alkalinized ropivacaine. In our experiment, low doses of bicarbonate produced significant increases in the proportion of nonionized ropivacaine with only modest precipitation. There would be a low likelihood of substantial drug precipitation if the mixture was administered within 5 to 10 minutes after alkalinization. These results indicate that alkalinized ropivacaine should not be used for infusions and that ropivacaine should not be alkalinized until just before use.

Amides↗

Laboratory measurements of oil mist concentrations using filters and an electrostatic precipitator.

This study investigated the potential for mineral oil mist to evaporate, during sampling, from filters and electrostatic precipitator substrates used to assess personal exposure. If sample evaporation occurs, reported mist concentrations will underestimate true exposure. Mineral oil used as a machining fluid is not normally considered volatile; however, when dispersed as mist its aggregate surface area is so high that significant evaporation can occur. National Institute for Occupational Safety and Health Method 5026 specifies that oil mist concentrations should be determined by collecting mist on filters of mixed cellulose esters (MCE) or polyvinyl chloride (PVC). Collected mist droplets remain dispersed on the filter surface and in contact with passing air while sampling continues, conditions that can lead to sample evaporation. Less evaporation should occur for samples taken with an electrostatic precipitator, where mist droplets are separated from the airflow by electrostatic force and coalesce on the precipitator wall to form a film with relatively low surface area. Collection of mineral oil mist was investigated using a precipitator designed for personal sampling and using either an MCE or a PVC filter. The amounts of oil mist collected using the precipitator were significantly higher than the amounts collected using the filters, p < 0.001. Further tests in which clean air passed through mist-loaded precipitators and filters showed that the precipitator retained substantially more collected mist than both filters, p < 0.001, and further suggested that the MCE filter retained more mist than the PVC filter, p = 0.059. Differences in sample collection and retention between the precipitator and the filters were particularly pronounced at mist loadings below 1 mg.

Aerosols↗

Calcium phosphate-containing precipitate and the carcinogenicity of sodium salts in rats.

Sodium saccharin, ascorbate and other sodium salts fed at high doses to rats produce urinary bladder urothelial cytotoxicity with consequent regenerative hyperplasia. For sodium salts that have been tested, tumor activity is enhanced when administered either alone or after a brief exposure to a known genotoxic bladder carcinogen. These sodium salts alter urinary composition of rats resulting in formation of an amorphous precipitate. We examined the precipitate to ascertain its composition and further delineate the basis for its formation in rat urine. Using scanning electron microscopy with attached X-ray energy dispersive spectroscopy, the principal elements present were calcium, phosphorus, minor amounts of silicon and sulfur. Smaller elements are not detectable by this method. Infrared analyses demonstrated that calcium phosphate was in the tribasic form and silicon was most likely in the form of silica. Small amounts of saccharin were present in the precipitate from rats fed sodium saccharin (<5%), but ascorbate was not detectable in the precipitate from rats fed similar doses of sodium ascorbate. Large amounts of urea and mucopolysaccharide, apparently chondroitin sulfate, were detected in the precipitate by infrared analysis. Chemical analyses confirmed the presence of large amounts of calcium phosphate with variably small amounts of magnesium, possibly present as magnesium ammonium phosphate crystals, present in urine even in controls. Small amounts of protein, including albumin and alpha(2u)-globulin, were also detected (<5% of the precipitate). Calcium phosphate is an essential ingredient of the medium for tissue culture of epithelial cells, but when present at high concentrations (>5 mM) it precipitates and becomes cytotoxic. The nature of the precipitate reflects the unique composition of rat urine and helps to explain the basis for the species specificity of the cytotoxic and proliferative effects of high doses of these sodium salts.

Animals↗